A rapid and efficient method for site-directed mutagenesis using one-step overlap extension PCR.

A Urban; S Neukirchen; K E Jaeger

doi:10.1093/nar/25.11.2227

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PubMed · 9153325

A rapid and efficient method for site-directed mutagenesis using one-step overlap extension PCR.

A Urban·S Neukirchen·K E Jaeger

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Abstract

A rapid method is described to efficiently perform site-directed mutagenesis based on overlap extension polymerase chain reaction (OE-PCR). Two template DNA molecules in different orientations relative to only one universal primer were amplified in parallel. By choosing a high dilution of mutagenic primers it was possible to run an overlap extension PCR in only one reaction without purification of intermediate products. This method which we have named one-step overlap extension PCR (OOE-PCR) can in principle be applied to every DNA fragment which can be cloned into a multiple cloning site of any common cloning vector.

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Animals DNA, Bacterial Deoxyribonucleases, Type II Site-Specific Green Fluorescent Proteins Lipase Luminescent Proteins Mutagenesis, Site-Directed Polymerase Chain Reaction Pseudomonas aeruginosa Scyphozoa Journal Article Research Support, Non-U.S. Gov't

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A Urban, S Neukirchen, K E Jaeger. 1997-06-01. A rapid and efficient method for site-directed mutagenesis using one-step overlap extension PCR.. https://doi.org/10.1093/nar%2F25.11.2227

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